Cationic lipid-mediated transfer of the hIL-10 gene prolongs survival of allogeneic hepatocytes in nagase analbuminemic rats

被引:11
|
作者
Fabrega, AJ
Fasbender, AJ
Struble, S
Zabner, J
机构
[1] UNIV IOWA,DEPT SURG,TRANSPLANTAT SECT,IOWA CITY,IA 52242
[2] UNIV IOWA,DEPT MED,IOWA CITY,IA 52242
[3] UNIV IOWA,HOWARD HUGHES MED INST,IOWA CITY,IA 52242
关键词
D O I
10.1097/00007890-199612270-00032
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Gene transfer techniques can be used as a drug delivery system to achieve local immunosuppression. We performed a series of experiments to identify the cationic lipid that most efficiently transfects isolated, cultured, rat hepatocytes; to optimize conditions for efficient transfection; to determine the duration of gene expression in vitro; and finally, to determine the survival of allogeneic hepatocytes transplanted into Nagase rats. Our results suggest that DOTAP is the best cationic lipid for transfection of cultured rat hepatocytes. In addition, the following conditions appear to optimize transfection efficiency: a DNA:DOTAP ratio of 1:6; a 24 exposure time of the hepatocytes to the DNA-DOTAP complex; a DNA dose of 4 mu g/35 mm culture plate seeded with 2.5 x 10(5) rat hepatocytes. When transfected as described above, cultured hepatocytes expressed the hIL-10 gene for approximately 14 days. Accordingly, Nagase rats transplanted with 4 x 10(7) DOTAP-hIL-10 transfected, allogeneic hepatocytes had an abrupt rise in serum albumin levels that peaked within 7 days of the transplant, decreased abruptly after 15 days, and approached baseline by day 40. In contrast, control animals had a smaller albumin peak. that returned to baseline within 10 days (P<0.01). In all animals, serum hIL-10 levels were undetectable when tested. We conclude that DOTAP is the best cationic lipid for transfection of cultured rat hepatocytes. Furthermore, hIL-10 transfected hepatocytes have a prolonged survival in an allogeneic horst which is probably limited by toss of gene expression. Further studies using other vectors capable of prolonged gene expression will help determine if indefinite hlL-10 gene expression leads to indefinite graft survival.
引用
收藏
页码:1866 / 1871
页数:6
相关论文
共 39 条
  • [21] Cationic lipid-mediated gene transfer: effect of serum on cellular uptake and intracellular fate of lipopolyamine/DNA complexes
    Escriou, V
    Ciolina, C
    Lacroix, F
    Byk, G
    Scherman, D
    Wils, P
    BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 1998, 1368 (02): : 276 - 288
  • [22] Lentivirus-mediated gene transfer of viral interleukin 10 prolongs survival of cardiac allografts
    Zhao, J
    Pettigrew, GJ
    Delriviere, L
    Bolton, EM
    Lever, AML
    Bradley, JA
    JOURNAL OF GENE MEDICINE, 2004, 6 (09): : S23 - S23
  • [23] Small poly-L-lysines improve cationic lipid-mediated gene transfer in vascular cells in vitro and in vivo
    Golda, Adam
    Pelisek, Jaroslav
    Klocke, Rainer
    Engelmann, Markus G.
    Rolland, Pierre-Henri
    Mekkaoui, Choukri
    Nikol, Sigrid
    JOURNAL OF VASCULAR RESEARCH, 2007, 44 (04) : 273 - 282
  • [24] The safety profile of a cationic lipid-mediated cystic fibrosis gene transfer agent following repeated monthly aerosol administration to sheep
    Alton, Eric W. F. W.
    Baker, Alison
    Baker, Eilidh
    Boyd, A. Christopher
    Cheng, Seng H.
    Coles, Rebecca L.
    Collie, D. David S.
    Davidson, Heather
    Davies, Jane C.
    Gill, Deborah R.
    Gordon, Catherine
    Griesenbach, Uta
    Higgins, Tracy
    Hyde, Stephen C.
    Innes, J. Alastair
    McCormick, Dominique
    McGovern, Michael
    McLachlan, Gerry
    Porteous, David J.
    Pringle, Ian
    Scheule, Ronald K.
    Shaw, Darren J.
    Smith, Sionagh
    Sumner-Jones, Stephanie G.
    Tennant, Peter
    Vrettou, Christina
    BIOMATERIALS, 2013, 34 (38) : 10267 - 10277
  • [25] Retrovirus-mediated transfer of viral IL-10 gene prolongs murine cardiac allograft survival
    Qin, LH
    Chavin, KD
    Ding, YZ
    Tahara, H
    Favaro, JP
    Woodward, JE
    Suzuki, T
    Robbins, PD
    Lotze, MT
    Bromberg, JS
    JOURNAL OF IMMUNOLOGY, 1996, 156 (06): : 2316 - 2323
  • [26] Synthetic DNA-compacting peptides derived from human sequence enhance cationic lipid-mediated gene transfer in vitro and in vivo
    B Schwartz
    M-A Ivanov
    B Pitard
    V Escriou
    R Rangara
    G Byk
    P Wils
    J Crouzet
    D Scherman
    Gene Therapy, 1999, 6 : 282 - 292
  • [27] Intratumoral accumulation and activation of natural killer cells following cationic lipid-mediated gene transfer of a human interleukin-2 transgene
    Clark, PR
    Parker, SE
    Hersh, EM
    CANCER GENE THERAPY, 1998, 5 (06) : S4 - S4
  • [28] Synthetic DNA-compacting peptides derived from human sequence enhance cationic lipid-mediated gene transfer in vitro and in vivo
    Schwartz, B
    Ivanov, MA
    Pitard, B
    Escriou, V
    Rangara, R
    Byk, G
    Wils, P
    Crouzet, J
    Scherman, D
    GENE THERAPY, 1999, 6 (02) : 282 - 292
  • [29] Adenovirus-mediated Bcl-2 gene transfer inhibits apoptosis and promotes survival of allogeneic transplanted hepatocytes
    Song, EW
    Chen, JS
    Antus, B
    Su, FX
    Wang, MH
    Exton, MS
    SURGERY, 2001, 130 (03) : 502 - 511
  • [30] Cationic lipid-mediated CFTR gene transfer to the lungs and nose of patients with cystic fibrosis:: a double-blind placebo-controlled trial
    Alton, EWFW
    Stern, M
    Farley, R
    Jaffe, A
    Chadwick, SL
    Phillips, J
    Davies, J
    Smith, SN
    Browning, J
    Davies, MG
    Hodson, ME
    Durham, SR
    Li, D
    Jeffery, PK
    Scallan, M
    Balfour, R
    Eastman, SJ
    Cheng, SH
    Smith, AE
    Meeker, D
    Geddes, DM
    LANCET, 1999, 353 (9157): : 947 - 954